Inflammatory mediators in alkali-burned corneas: preliminary characterization.
Inflammatory mediators in alkali-burned corneas: preliminary characterization.
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碱烧角膜中的炎症介质：初步表征。
作者：
S. A. Elgebaly;R. Downes;M. Bohr;F. Forouhar;J. O'Rourke;D. Kreutzer
白细胞趋化因子（LCF）是一种重要的炎症介质，可激活和募集循环中的白细胞进入组织损伤部位。最近在碱烧伤引起的炎症眼的泪液中检测到这些因子。然而，目前尚不清楚检测到的LCF是从受伤的角膜组织中释放出来的，还是从循环中泄漏出来的。使用我们实验室开发的角膜杯模型，我们开始检查角膜组织响应于碱损伤产生LCF的能力。我们还评估了柠檬酸对碱氢氧化钠（NaOH）预损伤角膜产生LCF的影响。对于这些研究，将从牛和人眼分离的角膜的上皮表面在室温下暴露于1 N NaOH 35秒。然后除去NaOH，用缓冲液洗涤上皮表面一次，并在37 ℃/5%CO2气氛下用培养基孵育1、2、4和6小时。结果表明：（1）NaOH_2可引起角膜上皮细胞损伤，损伤范围从上皮上半部的细胞变色和中度损伤（1-4小时）到上皮完全破坏（6小时）;（2）NaOH损伤的角膜（损伤后2小时孵育）产生显著水平的趋化活性（通过棋盘分析）特异于中性粒细胞（115%最大趋化反应[MCR]）和单核细胞（94% MCR）;（3）对这些因子的初步表征表明，它们是蛋白酶和热敏感的，可被有机溶剂提取，并且具有大于100，000道尔顿的分子量值;和（4）用0.01%柠檬酸孵育NaOH预处理的角膜2小时显著抑制嗜中性粒细胞（98%抑制）和单核细胞（91%抑制）的LCF产生。这些研究的结果表明，碱烧伤的牛和人角膜产生白细胞趋化因子，其生化特性不同于先前已知的低分子量趋化因子，如C5 a，白细胞介素-1，或白三烯B4。
Leukocyte chemotactic factors (LCF) are important inflammatory mediators which activate and recruit leukocytes from the circulation into sites of tissue damage. These factors were recently detected in the tear fluid of inflamed eyes induced by alkali burn. It remains unclear, however, whether the detected LCF are released from injured corneal tissues or leaked from the circulation. Using a corneal cup model developed in our laboratory, we began examining the capability of corneal tissues to produce LCF in response to alkali injury. We also evaluated the influence of citric acid on the production of LCF from corneas preinjured by the alkali sodium hydroxide (NaOH). For these studies, the epithelial surfaces of corneas isolated from bovine and human eyes were exposed to 1N NaOH for 35 seconds at room temperature. The NaOH was then removed and the epithelial surfaces washed once with buffer and incubated with culture medium for 1, 2, 4, and 6 hours at 37 degrees C/5% CO2 atmosphere. Our results showed that (1) NaOH2 induced corneal epithelial cell injury ranging from cell discoloration and moderate damage of the upper half of the epithelium (1-4 hrs) to total destruction of the epithelium (6 hrs); (2) NaOH-injured corneas (2 hr incubation post injury) produced significant levels of chemotactic activity (via checkerboard analysis) specific for neutrophils (115% maximum chemotactic response [MCR]) and mononuclear cells (94% MCR); (3) preliminary characterization of these factors revealed that they are protease and heat sensitive, extractable by organic solvents, and possess molecular weight values greater than 100,000 daltons; and (4) incubation of NaOH-pretreated corneas with 0.01% citric acid for 2 hours markedly inhibited the production of LCF for both neutrophils (98% inhibition) and mononuclear cells (91% inhibition). Results of these studies indicate that alkali-burned bovine and human corneas generate leukocyte chemoattractants which differ in their biochemical characteristics from previously known low molecular weight chemotactic factors such as C5a, interleukin-1, or leukotriene B4.
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DOI：
10.1172/jci111545
发表时间：
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期刊：
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影响因子：
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1984
期刊：
The American journal of pathology
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发表时间：
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影响因子：
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作者：
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